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22 July 2019 : Laboratory Research  

Neutrophil Gelatinase-Associated Lipocalin2 Exaggerates Cardiomyocyte Hypoxia Injury by Inhibiting Integrin β3 Signaling

Yang Gu1ABCDEF, Jin Geng1BCF, Zhuo Xu1CD, Yu Chen1DE, Xi-Wen Zhang1AG*

DOI: 10.12659/MSM.915108

Med Sci Monit 2019; 25:5426-5434

Abstract

BACKGROUND: The neutrophil inflammatory protein, lipocalin-2 (NGAL), is elevated in certain forms of cardiac hypertrophy and acute heart failure. However, the specific role of NGAL in cardiac hypoxia injury is unclear. This study aimed to elucidate the functional role of NGAL in cardiomyocyte hypoxia injury.

MATERIAL AND METHODS: Neonatal rat cardiomyocytes were transfected with adenovirus [(Ad-NGAL] to overexpress human-NGAL and then were exposed to hypoxia for 24 h to establish a hypoxia model. Cell inflammation was detected by RT-PCT and ELISA assay. Cell apoptosis was detected by TUNEL assay. Oxidative stress was also detected by commercial kits.

RESULTS: An increased inflammatory response, apoptosis, and augmented oxidative stress were observed after exposure to hypoxia, while NGAL overexpression in cells increased the expression and release of inflammatory cytokines. NGAL overexpression also increased the number of apoptotic cells and the imbalance of Bax/Bcl-2 protein expression. Moreover, NGAL overexpression increased the levels of reactive oxygen species and oxidase activity, but reduced anti-oxidase activity. Mechanistically, we found that NGAL decreased the expression of integrin β3, but not the expression of integrin avβ3 and avβ5, thus inhibiting the downstream protein AKT. When we used the constitutively activated AKT overexpression adenovirus to activate AKT, the deteriorated phenotype by NGAL was counteracted.

CONCLUSIONS: NGAL can directly affect cardiomyocytes and cause cardiomyocyte deteriorated hypoxia injury through inhibiting integrin β3 signaling.

Keywords: Cell Hypoxia, Integrin beta3, Myocytes, Cardiac, Acute-Phase Proteins, Animals, Newborn, Apoptosis, Cardiomegaly, Lipocalin-2, Lipocalins, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-bcl-2, Reactive Oxygen Species, Signal Transduction, bcl-2-Associated X Protein

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750